Bronchodilating beta-agonist compositions and methods

ABSTRACT

Bronchodilating compositions and methods are provided. The compositions are intended for administration as a nebulized aerosol. In certain embodiments, the compositions contain formoterol, or a derivative thereof. Methods for treatment, prevention, or amelioration of one or more symptoms of bronchoconstrictive disorders using the compositions provided herein are also provided.

RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.11/688,463, filed Mar. 20, 2007, which in turn is a continuation of U.S.patent application Ser. No. 10/887,785, filed Jul. 9, 2004, which, inturn claims priority under 35 U.S.C. §119(e) to U.S. Provisional PatentApplication No. 60/486,386, filed Jul. 10, 2003, entitled“BRONCHODILATING β-AGONIST COMPOSITIONS AND METHODS.” The disclosure ofeach of the above-referenced applications is incorporated by referenceherein in its entirety.

FIELD

Compositions and methods are provided relating to treatment, prevention,or amelioration of one or more symptoms of bronchoconstrictivedisorders. In particular, the compositions and methods herein includeformoterol, and/or derivatives thereof. The compositions arepropellant-free, sterile unit dose or multidose inhalation solutionsintended for administration via nebulization.

BACKGROUND

Bronchoconstrictive disorders affect millions worldwide. Such disordersinclude asthma (including bronchial asthma, allergic asthma andintrinsic asthma, e.g., late asthma and airway hyper-responsiveness),chronic bronchitis and other chronic obstructive pulmonary diseases.Compounds having β₂-adrenoreceptor agonist activity have been developedto treat these conditions. Such compounds include, but are not limitedto, Albuterol(a¹-(((1,1-dimethylethyl)amino)methyl)-4-hydroxy-1,3-benzenedimethanol);Bambuterol (dimethylcarbamic acid5-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,3-phenylene ester);Bitolterol (4-methylbenzoic acid4-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,2-phenylene ester);Broxaterol(3-bromo-a-(((1,1-methylethyl)amino)methyl)-5-isoxazolemethanol);Isoproterenol(4-(1-hydroxy-2-((1-methylethyl)amino)ethyl)-1,2-benzenediol);Trimetoquinol(1,2,3,4-tetrahydro-1-((3,4,5-trimethoxyphenyl)methyl)-6,7-isoquinolinediol);Clenbuterol(4-amino-3,5-dichloro-a-(((1,1-diemethylethyl)amino)-methyl)benzenemethanol);Fenoterol(5-(1-hydroxy-2-((2-(4-hydroxyphenyl)-1-methylethyl)-amino)ethyl)-1,3-benzenediol);Formoterol(2-hydroxy-5-((1RS)-1-hydroxy-2-(((1RS)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide);(R,R)-Formoterol; Desformoterol ((R,R) or(S,S)-3-amino-4-hydroxy-a-(((2-(4-methoxyphenyl)-1-ethylethyl)amino)methyl)-benzene-methanol);Hexoprenaline(4,4′-(1,6-hexanediyl)-bis(imino-(1-hydroxy-2,1-ethane-diyl)))bis-1,2-benzenediol);Isoetharine(4-(1-hydroxy-2-((1-methylethyl)amino)butyl)-1,2-benzenediol);Isoprenaline(4-(1-hydroxy-2-((1-methylethyl)amino)ethyl)-1,2-benzenediol);Metaproterenol(5-(1-hydroxy-2-((1-methylethyl)amino)ethyl)-1,3-benzenediol);Picumeterol(4-amino-3,5-dichloro-a-(((6-(2-(2-pyridinyl)ethoxy)hexyl)-amino)methyl)benzenemetlianol);Pirbuterol(a⁶-(((1,1-dimethylethyl)amino)methyl)-3-hydroxy-2,6-pyridinemethanol);Procaterol (((R*,S*)-(±)-8-hydroxy-5-(1-hydroxy-2-((1-methylethyl)amino)butyl)-2-(1H)-quinolinone); Reproterol((7-(3-((2-(3,5-dihydroxy-phenyl)-2-hydroxyethyl)amino)propyl)-3,7-dihydro-1,3-dimethyl-1H-purine-2,6-dione);Rimiterol (4-(hydroxy-2-piperidinylmethyl)-1,2-benzenediol); Salbutamol((±)-a¹-(((1,1-dimethylethyl)amino)methyl)-4-hydroxy-1,3-benzenedimethanol);(R)-Salbutamol; Salmeterol((±)-4-hydroxy-a¹-(((6-(4-phenylbutoxy)hexyl)amino)methyl)-1,3-benzenedimethanol);(R)-Salmeterol; Terbutaline(5-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,3-benzenediol);Tulobuterol(2-chloro-a-(((1,1-dimethyl-ethyl)amino)-methyl)benzenemethanol); andTA-2005(8-hydroxy-5-((1R)-1-hydroxy-2-(N-((1R)-2-(4-methoxyphenyl)-1-methylethyl)amino)ethyl)carbostyrilhydrochloride).

These compounds are typically formulated for inhalation therapy. Aqueousor liquid formulations are preferred over solid formulations. Powderedformulations are more difficult to administer, particularly to the youngand elderly who are most often the patients in need of such therapy.Compounds, such as formoterol are not adequately stable in aqueoussolutions to be formulated as liquids. Hence there is a need forformulations of compounds, such as formoterol, in a form that can beconveniently administered and that are stable for extended periods oftime.

SUMMARY

Compositions and methods for treatment, prevention, or amelioration ofone or more symptoms of bronchoconstrictive disorders are provided. Thecompositions provided herein are stable solutions of a bronchodilatingagent, or a derivative thereof, in a pharmacologically suitable fluidthat contains water, that are stable during long term storage. Thecompositions are suitable for direct administration to a subject in needthereof. Pharmacologically suitable fluids include, but are not limitedto, polar fluids, including protic fluids. In certain embodimentsherein, the compositions are aqueous solutions.

The compositions provided herein possess an estimated shelf-life ofgreater than 1, 2 or 3 months usage time at 25° C. and greater than orequal to 1, 2 or 3 years storage time at 5° C. In certain of theseembodiments, using Arrhenius kinetics, >80% or >85% or >90% or >95%estimated bronchodilating agent remains after such storage. Thesecompositions are particularly useful for administration vianebulization. In certain embodiments herein, the subject is a mammal. Inother embodiments, the subject is a human.

The compositions provided herein are formulated to remain stable over arelatively long period of time. For example, the compositions providedherein are stored between −15° C. and 25° C., or between 2° C. and 8°C., and remain stable for the desired time. In one embodiment, thecompositions are stored at 5° C. In other embodiment, the compositionsare stored at 25° C.

Among the bronchodilating agents for use herein are Albuterol(a¹-(((1,1-dimethylethyl)-amino)methyl)4-hydroxy-1,3-benzenedimethanol);Bambuterol (dimethylcarbamic acid5-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,3-phenylene ester);Bitolterol (4-methylbenzoic acid4-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,2-phenylene ester);Broxaterol(3-bromo-a-(((1,1-dimethylethyl)amino)-methyl)-5-isoxazolemethanol);Isoproterenol(4-(1-hydroxy-2-((1-methylethyl)amino)ethyl)-1,2-benzenediol);Trimetoquinol(1,2,3,4-tetrahydro-1-((3,4,5-trimethoxyphenyl)methyl)-6,7-isoquinolinediol);Clenbuterol(4-amino-3,5-dichloro-a-(((1,1-dimethylethyl)amino)methyl)benzenemethanol);Fenoterol(5-(1-hydroxy-2-((2-(4-hydroxyphenyl)-1-methylethyl)amino)ethyl)-1,3-benzenediol);Formoterol(2-hydroxy-5-((1RS)-1-hydroxy-2-(((1RS)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)-formanilide);(R,R)-Formoterol; (S,S)-Formoterol; Desformoterol ((R,R) or(S,S)-3-amino-4-hydroxy-a-(((2-(4-methoxyphenyl)-1-methylethyl)amino)methyl)benzene-methanol);Hexoprenaline(4,4′-(1,6-hexanediyl)-bis(imino(1-hydroxy-2,1-ethane-diyl)-))bis-1,2-benzenediol);Isoetharine(4-(1-hydroxy-2-((1-methylethyl)amino)-butyl)-1,2-benzenediol);Isoprenaline(4-(1-hydroxy-2-((1-methylethyl)amino)ethyl)-1,2-benzenediol);Metaproterenol(5-(1-hydroxy-2-((1-methylethyl)-amino)ethyl)-1,3-benzenediol);Picumeterol(4-amino-3,5-dichloro-a-(((6-(2-(2-pyridinyl)-ethoxy)hexyl)-amino)methyl)benzenemethanol);Pirbuterol(a⁶-(((1,1-dimethylethyl)amino)-methyl)-3-hydroxy-2,6-pyridinemethanol);Procaterol(((R*,S*)-(±)-8-hydroxy-5-(1-hydroxy-2-((1-methylethyl)amino)butyl)-2(1H)-quinolinone);Reproterol((7-(3-((2-(3,5-dihydroxyphenyl)-2-hydroxyethyl)amino)propyl)-3,7-dihydro-1,3-dimethyl-1H-purine-2,6-dione);Rimiterol (4-(hydroxy-2-piperidinylmethyl)-1,2-benzenediol); Salbutamol((±)-a¹-(((1,1-dimethylethyl)-amino)methyl)-4-hydroxy-1,3-benzenedimethanol);(R)-Salbutamol; Salmeterol((±)-4-hydroxy-a¹-(((6-(4-phenylbutoxy)hexyl)amino)methyl)-1,3-benzenedimethanol);(R)-Salmeterol; Terbutaline(5-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,3-benzenediol);Tulobuterol(2-chloro-a-(((1,1-dimethyl-ethyl)amino)methyl)benzene-methanol); andTA-2005(8-hydroxy-5-((1R)-1-hydroxy-2-(N-((1R)-2-(4-methoxy-phenyl-1-methylethyl)amino)ethyl)carbostyrilhydrochloride).

Of particular interest herein is formoterol, having the formula:

Formoterol for use in the compositions and methods provided hereinincludes2-hydroxy-5-((1RS)-1-hydroxy-2-(((1RS)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide;or a stereoisomer thereof; and also includes the single enantiomers2-hydroxy-5-((1S)-1-hydroxy-2-(((1S)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilideand2-hydroxy-5-((1R)-1-hydroxy-2-(((1R)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide.

In certain embodiments, the compositions are administered vianebulization. Administration of a nebulized aerosol is preferred overthe use of dry powders for inhalation in certain subject populations,including pediatric and geriatric groups.

In one embodiment, the compositions for use in the methods providedherein contain a pharmaceutically acceptable derivative of formoterol.In another embodiment, the compositions for use in the methods providedherein contain a pharmaceutically acceptable salt of formoterol.Pharmaceutically acceptable salts include, but are not limited to, saltsof mineral acids, such as but not limited to hydrochlorides andsulfates; and salts of organic acids, such as but not limited toacetates, lactates, malates, tartrates, citrates, ascorbates,succinates, butyrates, valerates and fumarates. In one embodiment, thecompositions for use in the methods provided herein contain formoterolfumarate or formoterol fumarate dihydrate. In another embodiment, thecompositions for use in the methods provided herein contain formoteroltartrate.

Also provided herein are combinations containing a composition providedherein and a nebulizer. The combinations can be packaged as kits, whichoptionally contain other components, including instructions for use ofthe nebulizer. Any nebulizer is contemplated for use in the kits andmethods provided herein. In particular, the nebulizers for use hereinnebulize liquid formulations, including the compositions providedherein, containing no propellant. The nebulizer may produce thenebulized mist by any method known to those of skill in the art,including, but not limited to, compressed air, ultrasonic waves, orvibration. The nebulizer may further have an internal baffle. Theinternal baffle, together with the housing of the nebulizer, selectivelyremoves large droplets from the mist by impaction and allows thedroplets to return to the reservoir. The fine aerosol droplets thusproduced are entrained into the lung by the inhaling air/oxygen.

Methods for the treatment, prevention, or amelioration of one or moresymptoms of bronchoconstrictive disorders, including, but not limitedto, asthma, including, but not limited to, bronchial asthma, allergicasthma and intrinsic asthma, e.g., late asthma and airwayhyper-responsiveness; chronic bronchitis; and other chronic obstructivepulmonary diseases are provided. The methods involve administering aneffective amount of a pharmaceutical composition provided herein to asubject in need of such treatment.

Articles of manufacture, containing packaging material, a compositionprovided herein, which is useful for treatment, prevention oramelioration of one or more symptoms of diseases or disorders associatedwith undesired and/or uncontrolled bronchoconstriction, and a label thatindicates that the composition is used for treatment, prevention oramelioration of one or more symptoms of diseases or disorders associatedwith undesired and/or uncontrolled bronchoconstriction, are alsoprovided.

DETAILED DESCRIPTION Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as is commonly understood by one of ordinary skillin the art to which this invention belongs. All patents, applications,published applications and other publications are incorporated byreference in their entirety. In the event that there are a plurality ofdefinitions for a term herein, those in this section prevail unlessstated otherwise.

As used herein, formoterol refers to2-hydroxy-5-((1RS)-1-hydroxy-2-(((1RS)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide;or a stereoisomer thereof. The term formoterol also refers to the singleenantiomers2-hydroxy-5-((1S)-1-hydroxy-2-(((1S)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide((S,S)-formoterol) and2-hydroxy-5-((1R)-1-hydroxy-2-(((1R)-2-(p-methoxyphenyl)-1-methylethyl)amino)-ethyl)formanilide((R,R)-formoterol).

As used herein, formoterol fumarate refers to a salt of formoterolhaving the formula (formoterol)·½ fumarate.

As used herein, formoterol free base refers to the neutral, anhydrousform of formoterol. Thus, a recitation that a composition contains,e.g., 20 μg/mL of formoterol free base means that the compositioncontains 20 μg/mL of neutral, anhydrous formoterol. Such compositionsmay be prepared using a derivative of formoterol.

As used herein, an aerosol is liquid or particulate matter dispersed inair. Aerosols include dispersions of liquids, including aqueous andother solutions, and solids, including powders, in air.

As used herein, a nebulized solution refers to a solution that isdispersed in air to form an aerosol. Thus, a nebulized solution is aparticular form of an aerosol.

As used herein, a nebulizer is an instrument that is capable ofgenerating very fine liquid droplets for inhalation into the lung.Within this instrument, the nebulizing liquid or solution is atomizedinto a mist of droplets with a broad size distribution by methods knownto those of skill in the art, including, but not limited to, compressedair, ultrasonic waves, or a vibrating orifice. Nebulizers may furthercontain, e.g., a baffle which, along with the housing of the instrument,selectively removes large droplets from the mist by impaction. Thus, themist inhaled into the lung contains fine aerosol droplets.

As used herein, a pharmacologically suitable fluid is a solvent suitablefor pharmaceutical use which is not a liquified propellant gas.Exemplary pharmacologically suitable fluids include polar fluids,including protic fluids such as water.

As used herein, a kit refers to one or more items, including, but notlimited to, compounds, compositions, combinations, instruments anddevices, suitably packaged for use. Kits provided herein optionallycontain instructions for use.

As used herein, a combination refers to any association between two oramong more items.

As used herein, fluid refers to any composition that can flow. Fluidsthus encompass compositions that are in the form of semi-solids, pastes,solutions, aqueous mixtures, gels, lotions, creams and other suchcompositions.

As used herein, a mixture is a mutual incorporation of two or moresubstances, without chemical union, the physical characteristics of eachof the components being retained.

As used herein, the stability of a composition provided herein refers tothe length of time at a given temperature that is greater than 80%, 85%,90% or 95% of the initial amount of active ingredient, e.g., formoterol,is present in the composition. Thus, for example, a composition that isstable for 30 days at 25° C. would have greater than 80%, 85%, 90% or95% of the initial amount of active ingredient present in thecomposition at 30 days following storage at 25° C.

As used herein, pharmaceutically acceptable derivatives of a compoundinclude salts, esters, enol ethers, enol esters, acids, bases, solvates,hydrates or prodrugs thereof. Such derivatives may be readily preparedby those of skill in this art using known methods for suchderivatization. The compounds produced may be administered to animals orhumans without substantial toxic effects and either are pharmaceuticallyactive or are prodrugs. Pharmaceutically acceptable salts include, butare not limited to, amine salts, such as but not limited toN,N′-dibenzylethylediamine, chloroprocaine, choline, ammonia,diethanolamine and other hydroxyalkylamines, ethylenediamine,N-methylglucamine, procaine, N-benzylphenethylamine,1-para-chlorobenzyl-2-pyrrolidin-1′-ylmethylbenzimidazole, diethylamineand other alkylamines, piperazine and tris(hydroxymethyl)aminomethane;alkali metal salts, such as but not limited to lithium, potassium andsodium; alkali earth metal salts, such as but not limited to barium,calcium and magnesium; transition metal salts, such as but not limitedto zinc; and other metal salts, such as but not limited to sodiumhydrogen phosphate and disodium phosphate; and also including, but notlimited to, salts of mineral acids, such as but not limited tohydrochlorides and sulfates; and salts of organic acids, such as but notlimited to acetates, lactates, malates, tartrates, citrates, ascorbates,succinates, butyrates, valerates and fumarates. Pharmaceuticallyacceptable esters include, but are not limited to, alkyl, alkenyl,alkynyl, aryl, heteroaryl, aralkyl, heteroaralkyl, cycloalkyl andheterocyclyl esters of acidic groups, including, but not limited to,carboxylic acids, phosphoric acids, phosphinic acids, sulfonic acids,sulfinic acids and boronic acids. Pharmaceutically acceptable enolethers include, but are not limited to, derivatives of formula C═C(OR)where R is hydrogen, alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl,heteroaralkyl, cycloalkyl and heterocyclyl. Pharmaceutically acceptableenol esters include, but are not limited to, derivatives of formulaC═C(OC(O)R) where R is hydrogen, alkyl, alkenyl, alkynyl, aryl,heteroaryl, aralkyl, heteroaralkyl, cycloalkyl and heterocyclyl.Pharmaceutically acceptable solvates and hydrates are complexes of acompound with one or more solvent or water molecule, in certainembodiments 1 to about 100, in other embodiments 1 to about 10, infurther embodiments one to about 2, 3 or 4, solvent or water molecules.Formoterol salts and hydrates are used in certain embodiments herein.

As used herein, treatment means any manner in which one or more of thesymptoms of a condition, disorder or disease are ameliorated orotherwise beneficially altered. Treatment also encompasses anypharmaceutical use of the compositions herein, such as use for treatingcancer.

As used herein, amelioration of the symptoms of a particular disorder byadministration of a particular pharmaceutical composition refers to anylessening, whether permanent or temporary, lasting or transient that canbe attributed to or associated with administration of the composition.

As used herein, a prodrug is a compound that, upon in vivoadministration, is metabolized or otherwise converted to thebiologically, pharmaceutically or therapeutically active form of thecompound. To produce a prodrug, the pharmaceutically active compound ismodified such that the active compound will be regenerated by metabolicprocesses. The prodrug may be designed to alter the metabolic stabilityor the transport characteristics of a drug, to mask side effects ortoxicity, to improve the flavor of a drug or to alter othercharacteristics or properties of a drug. By virtue of knowledge ofpharmacodynamic processes and drug metabolism in vivo, those of skill inthis art, once a pharmaceutically active compound is known, can designprodrugs of the compound (see, e.g., Nogrady (1985) Medicinal ChemistryA Biochemical Approach, Oxford University Press, New York, pages388-392).

It is to be understood that the compounds for use in the compositionsand methods provided herein may contain chiral centers. Such chiralcenters may be of either the (R) or (S) configuration, or may be amixture thereof. Thus, the compounds for use in the compositionsprovided herein may be enantiomerically pure, or be stereoisomeric ordiastereomeric mixtures. It is to be understood that the chiral centersof the compounds provided herein may undergo epimerization in vivo.Thus, one of skill in the art will recognize that administration of acompound in its (R) form is equivalent, for compounds that undergoepimerization in vivo, to administration of the compound in its (S)form.

As used herein, bronchoconstriction refers to a reduction in the caliberof a bronchus or bronchi.

As used herein, undesired and/or uncontrolled bronchoconstriction refersto bronchoconstriction that results in or from a pathological symptom orcondition. Pathological conditions include, but are not limited to,asthma and chronic obstructive pulmonary disease (COPD). Pathologicalsymptoms include, but are not limited to, asthma and COPD.

As used herein, the statement that a composition is stable during “longterm storage” means that the composition is suitable for administrationto a subject in need thereof when it has an estimated shelf-life ofgreater than 1, 2 or 3 months usage time at 25° C. and greater than orequal to 1, 2 or 3 years storage time at 5° C. In certain embodimentsherein, using Arrhenius kinetics, >80% or >85% or >90% or >95% estimatedbronchodilating agent remains after such storage.

A. Formoterol

Formoterol(2-hydroxy-5-((1RS)-1-hydroxy-2-(((1RS)-2-(p-methoxyphen-yl)-1-methylethyl)amino)ethyl)formanilide)is derived from adrenaline and, as noted above, is used as aβ₂-stimulator in inhalation therapy of respiratory diseases,particularly for the treatment of bronchial asthma. It has been reportedthat in patients with reversible obstructive respiratory diseases,formoterol has a bronchodilatory effect. This effect has a relativelyrapid onset (approximately 1-3 minutes) and a relatively long duration(greater than 12 hours). Formoterol inhibits the release of leukotrienesand other messenger substances involved with inflammation, such ashistamines. In addition, formoterol may bring about a hyperglycemicactivity.

To date, formoterol has been formulated as a dry powder and administeredvia devices such as the TURBUHALER® and the AEROLIZER®. See, e.g.,Seberova et al. (2000) Respir. Med. 94(6):607-6111; Lotvall et al.(1999) Can. Respir. J. 6(5):412-416; Campbell et al. (1999) Respir. Med.93(4):236-244; Nightingale et al. (1999) Am. J. Respir. Crit. Care Med.159(6):1786-1790; Lecaillon et al. (1999) Eur. J. Clin. Pharmacol.55(2):131-138; Bartow et al. (1998) Drugs 55(2):303-322; Ekstrom et al.(1998) Respir. Med. 92(8): 1040-1045; Ringdal et al. (1998) Respir. Med.92(8):1017-1021; Totterman et al. (1998) Eur. Respir. J. 12(3):573-579;Palmqvist et al. (1997) Eur. Respir. J. 10(11):2484-2489; Nielsen et al.(1997) Eur. Respir. J. 10(9):2105-2109; Ullman et al. (1996) Allergy51(10):745-748; Seiroos et al. (1996) Clin. Immunother. 6:273-299; andSchreurs et al. (1996) Eur. Respir. J. 9(8):1678-1683.

Formoterol is also available as a tablet and a dry syrup in certainareas of the world (e.g., ATOCK®, marketed by Yamanouchi PharmaceuticalCo. Ltd., Japan). Formoterol formulations are also available in otherareas (e.g., Europe and U.S.) for propellant-based metered dose inhalersand dry powder inhalers (e.g., TURBUHALER®, AEROLIZER® AND FORADILAEROLIZER®). None of these formulations are water based. Sterile,stable, aqueous based inhalation solutions of formoterol fornebulization are not available, nor have they been reported.

In the treatment of bronchoconstrictive diseases, sufficient amount ofthe inhaled drug should reach their local site of action in order to beefficacious. It is known that different delivery methods and deliverydevices have different deposition characteristics. Consequently, underoptimal inhalation conditions, doses from different delivery methods anddelivery devices result in different delivered doses and differentamounts deposited at the active site. The actual dose reaching theactive site also depends upon the amount of drug particles included inthe delivered dose and the inhalation characterstics of the patient. Nocorrelation between the amount of drug administered by dry powderinhalers (DPIs) or metered dose inhalers (MDIs) and the actual amountthat gets deposited at the active site has been established so far. Norhas a correlation been established between DPI or MDI dosages andnebulization dosages.

Compositions containing formoterol in combination with other activeingredients have been disclosed. See, e.g., U.S. Pat. Nos. 6,004,537,5,972,919 and 5,674,860 (formoterol and budenoside), U.S. Pat. Nos.5,668,110, 5,683,983, 5,677,280 and 5,654,276 (formoterol and IL-5inhibitors), U.S. Pat. No. 6,136,603 (formoterol and antisensemodulators of IL-5), U.S. Pat. No. 5,602,110 (formoterol andmillrinone), U.S. Pat. No. 5,525,623 (formoterol and a tryptaseinhibitor), U.S. Pat. Nos. 5,691,336, 5,877,191, 5,929,094, 5,750,549and 5,780,467 (formoterol and a tachykinin receptor antagonist); andInternational Patent Application Publication Nos. WO 99/00134(formoterol and rofleponide) and WO 99/36095 (formoterol and a dopamineD₂ receptor agonist).

Other compositions containing formoterol have been disclosed in U.S.Pat. Nos. 5,677,809, 6,126,919, 5,733,526, 6,071,971, 6,068,833,5,795,564, 6,040,344, 6,041,777, 5,874,481, 5,965,622 and 6,161,536.

U.S. Pat. No. 6,150,418 discloses a “liquid active substanceconcentrate” containing formoterol in the form of its free base or inthe form of one of the pharmacologically acceptable salts or additionproducts (adducts) thereof as active substance. This “liquid activesubstance concentrate” is reported to be a concentrated (i.e., greaterthan 10 mg/ml, preferably 75 to 500 mg/mL) solution or suspension thatis stable for a period of several months possibly up to several yearswithout any deterioration in the pharmaceutical quality. This patentteaches that it is the high concentration that allows for the stabilityof the concentrate. The “liquid active substance concentrate” is notsuitable for direct administration to a patient.

U.S. Pat. No. 6,040,344 discloses an aqueous aerosol formulation offormoterol tartrate for use in a nebulizer. This patent states that theformulation disclosed therein is not attractive for long term storage.

B. Compositions for Use in Treatment, Prevention, or Amelioration of Oneor More Symptoms of Bronchoconstrictive Disorders

Pharmaceutical compositions containing a β₂-adrenoreceptor agonist foradministration via nebulization are provided. The compositions aresterile filtered and filled in vials, including unit dose vialsproviding sterile unit dose formulations which are used in a nebulizerand suitably nebulized. Each unit dose vial is sterile and is suitablynebulized without contaminating other vials or the next dose.

The unit dose vials are formed in a form-fill-seal machine or by anyother suitable method known to those of skill in the art. The vials maybe made of plastic materials that are suitably used in these processes.For example, plastic materials for preparing the unit dose vialsinclude, but are not limited to, low density polyethylene, high densitypolyethylene, polypropylene and polyesters. In one embodiment, theplastic material is low density polyethylene.

In one embodiment, the β₂-adrenoreceptor agonist is formoterol, or apharmaceutically acceptable derivative thereof. In other embodiments,the formoterol for use in the compositions provided herein is formoterolfumarate. Formoterol refers to2-hydroxy-5-((1RS)-1-hydroxy-2-(((1RS)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide;or a stereoisomer thereof. The term formoterol also refers herein to thesingle enantiomers2-hydroxy-5-((1S)-1-hydroxy-2-(((1S)-2-p-methoxypheny-1)-1-methylethyl)amino)ethyl)formanilideand2-hydroxy-5-((1R)-1-hydroxy-2-(((1R)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide.

In certain embodiments, the compositions contain formoterol fumarate ata concentration of about 0.1 μg/mL up to about 150 μg/mL, or 0.1 μg/mLup to 150 μg/mL. In further embodiments, the compositions containformoterol fumarate at a concentration of about 0.1 μg/mL up to about100 μg/mL, or 0.11 g/mL up to 100 μg/mL. The formoterol fumarate isformulated, in certain compositions provided herein, at a concentrationof about 0.1 μg/mL up to 50 μg/mL, or 0.1 μg/mL up to 50 μg/m L. Infurther embodiments, the compositions contain formoterol fumarate at aconcentration of about 0.1 μg/mL up to about 40 μg/mL, or 0.1 μg/mL upto 40 μg/mL. In further embodiments, the compositions contain formoterolfumarate at a concentration of about 0.1 μg/mL up to about 20 μg/mL, or0.1 μg/mL up to 20 μg/mL. The formoterol fumarate is formulated, inother compositions provided herein, at a concentration of about 40μg/mL, or 40 μg/mL. In further embodiments, the compositions containformoterol fumarate at a concentration of about 35 μg/mL, or 35 μg/mL.In other embodiments, the compositions contain formoterol fumarate at aconcentration of about 30 μg/mL, or 30 μg/mL. In other embodiments, thecompositions contain formoterol fumarate at a concentration of about 25μg/mL, or 25 μg/L. In further embodiments, the compositions containformoterol fumarate at a concentration of about 20 μg/mL, or 20 μg/mL.In another embodiment, the compositions contain formoterol fumarate at aconcentration of about 15 μg/mL, or 15 μmL. In another embodiment, thecompositions contain formoterol fumarate at a concentration of about 12μg/mL, or 12 μg/mL. In another embodiment, the compositions containformoterol fumarate at a concentration of about 10 μg/mL, or 10 μg/mL.In another embodiment, the compositions contain formoterol fumarate at aconcentration of about 8 μg/mL, or 8 μg/mL. In another embodiment, thecompositions contain formoterol fumarate at a concentration of about 5μg/mL, or 5 μg/mL. In another embodiment, the compositions containformoterol fumarate at a concentration of about 2.5 μg/mL, or 2.5 μg/mL.In another embodiment, the compositions contain formoterol fumarate at aconcentration of about 1 μg/mL, or 1 μg/mL.

In certain embodiments, the compositions contain formoterol free base ata concentration of about 0.08 μg/mL up to about 128 μg/mL, or 0.08 μg/mLup to 128 μg/mL. In further embodiments, the compositions containformoterol free base at a concentration of about 0.08 μg/mL up to about86 μg/mL, or 0.08 μg/mL up to 86 μg/mL. The formoterol free base isformulated, in certain compositions provided herein, at a concentrationof about 0.08 μg/mL up to 43 μg/mL, or 0.08 μg/mL up to 43 μg/mL. Infurther embodiments, the compositions contain formoterol free base at aconcentration of about 0.08 μg/mL up to about 34 μg/mL, or 0.08 μg/mL upto 34 μg/mL. In further embodiments, the compositions contain formoterolfree base at a concentration of about 0.08 μg/mL up to about 26 μg/mL,or 0.08 μg/mL up to 26 μg/mL. The formoterol free base is formulated, inother compositions provided herein, at a concentration of about 0.08μg/ml, up to about 17 μg/mL, or 0.08 μg/mL up to 17 μg/mL. In furtherembodiments, the compositions contain formoterol free base at aconcentration of about 34 μg/mL, or 34 μg/mL. In further embodiments,the compositions contain formoterol free base at a concentration ofabout 30 μg/mL, or 30 μg/mL. In other embodiments, the compositionscontain formoterol free base at a concentration of about 25.6 μg/mL, or25.6 μg/mL. In further embodiments, the compositions contain formoterolfree base at a concentration of about 21.4 μg/mL, or 21.4 μg/mL. Infurther embodiments, the compositions contain formoterol free base at aconcentration of about 17 μg/mL, or 17 μg/mL. In another embodiment, thecompositions contain formoterol free base at a concentration of about 13μg/mL, or 13 μg/mL. In another embodiment, the compositions containformoterol free base at a concentration of about 10 μg/mL, or 10 μg/mL.In another embodiment, the compositions contain formoterol free base ata concentration of about 9 μg/mL, or 9 μg/mL. In another embodiment, thecompositions contain formoterol free base at a concentration of about 7μg/mL, or 7 μg/mL. In another embodiment, the compositions containformoterol free base at a concentration of about 4 μg/mL, or 4 μg/mL. Inanother embodiment, the compositions contain formoterol free base at aconcentration of about 2 μg/mL, or 2 μg/mL. In another embodiment, thecompositions contain formoterol free base at a concentration of about0.8 μg/mL, or 0.8 μg/mL.

The volume of formoterol inhalation solution nebulized depends on thenebulizer used. In certain embodiments, the volume is from about 0.1 mLup to about 3 mL, or 0.1 mL up to 3 mL. In other embodiments, the volumeis about 2 mL, or 2 mL. In other embodiments, the volume is about 1 mL,or 1 mL. In other embodiments, the volume is about 0.5 mL, or 0.5 mL.

The compositions containing the β₂-adrenoreceptor agonist, includingformoterol, are formulated with a pharmacologically suitable fluidPharmacologically suitable fluids include, but are not limited to, polarsolvents, including, but not limited to, compounds that contain hydroxylgroups or other polar groups. Such solvents include, but are not limitedto, water or alcohols, such as ethanol, isopropanol, and glycolsincluding propylene glycol, polyethylene glycol, polypropylene glycol,glycol ether, glycerol and polyoxyethylene alcohols.

Polar solvents also include protic solvents, including, but not limitedto, water, aqueous saline solutions with one or more pharmaceuticallyacceptable salt(s), alcohols, glycols or a mixture thereof. For a salinesolution as the solvent or as a component thereof, particularly suitablesalts are those which display no or only negligible pharmacologicalactivity after administration.

In the embodiments herein, the compositions have a pH of about 2.0 toabout 8.0, or 2.0 to 8.0. Ea other embodiments, the compositions have apH of about 4.0 to about 6.0, or 4.0 to 6.0. In other embodiments, thepH is about 4.5 to about 5.5, or 4.5 to 5.5. In certain of the aboveembodiments, the compositions are formulated at a pH of about 4, 4.4 or4.6 up to about 5.5, 5.7 or 6; or 4, 4.4 or 4.6 up to 5.5, 5.7 or 6. Inother embodiments, the pH is about 5.0, or 5.0. It has been found thatthe rate constant for decomposition of an aqueous solution of formoterolis dependent on pH. The rate constant (k_(obs)) at 60° C. at a pH of 3,4, 5 and 7 is approximately 0.62, 0.11, 0.044 and 0.55 day.sup.-1,respectively. Therefore, the decomposition of formoterol in aqueoussolution at 60° C. at a buffer concentration of 5 mM and an ionicstrength of 0.05 is slowest at a pH of about 5.0, or 5.0.

The solubility of formoterol in aqueous solution has been found to bedependent on pH. Thus, at a pH of between about 5 and about 7, theaqueous solubility of formoterol at ambient temperature is approximately2.2 mg/mL. At a pH of about 4, the aqueous solubility of formoterol atambient temperature is approximately 3 mg/mL, while at a pH of about 3,the aqueous solubility of formoterol at ambient temperature is about 4.8mg/mL. The solubility of formoterol in pure water, for example, highperformance liquid chromatography (HPLC) water, at ambient temperatureis approximately 2 mg/mL.

In other of the above embodiments, the compositions ether contain abuffer, including, but not limited to, citric acid/phosphate, acetate,barbital, borate, Britton-Robinson, cacodylate, citrate, collidine,formate, maleate, McIlvaine, phFosphate, Prideaux-Ward, succinate,citrate-phosphate-borate (Teorell-Stanhagen), veronal acetate, MES(2-(N-morpholino)ethanesulfonic acid), BIS-TRIS(bis(2-hydroxyethyl)imino-tris-(hydroxymethyl)methane), ADA(N-(2-acetamido)-2-iminodiacetic acid), ACES(N-(carbamoylmethyl)-2-aminoethanesulfonic acid), PIPES(piperazine-N,N′-bis(2-ethanesulfonic acid)), MOPSO(3-(N-morpholino)-2-hydroxypropanesulfonic acid), BISTRIS PROPANE(1,3-bis(trisfhydroxymethyl)methylamino)propane), BES(N,N-bis(2-hydroxyethyl)-2-arainoethanesulfonic acid), MOPS(3-N-morpholinopropanesulfonic acid), TES(N-tris(hydroxymethyl)methyl—2-aminoethanesulfonic acid), HEPES(N-(2-hydroxyethyl)-piperazine-N′-(2-ethanesulfonic acid), DIPSO(3-(N,N-bis(2-hydroxyethyl)amino)-2-hydroxypropanesulfonic acid), MOBS(4-(N-morpholino)-butanesulfonic acid), TAPSO(3-(N-tris(hydroxymethyl)methylamino)-2-hydroxypropanesulfonic acid),TRIZMA® (tris(hydroxymethylaminomethane),HEPPSO(N-(2-hydroxyethyl)piperazine-N′-(2-hydroxy-propanesulfonic acid),POPSO (piperazine-N,N′-bis(2-hydroxypropanesulfonic acid)), TEA(triethanolamine),EPPS(N-(2-hydroxyethyl)piperazine-N′-(3-propanesulfonic acid), TRICINE(N-tris(hydroxy-methyl)methylglycine), GLY-GLY (glycylglycine), BICINE(N,N-bis(2-hydroxyethyl)glycine), HEPBS(N-(2-hydroxyethyl)piperazine-N′-(4-butanesulfonic acid)),TAPS(N-trisfhydroxymethyl)methyl-3-aminopropanesulfonic acid), AMPD(2-amino-2-methyl-1,3-propanediol), and/or any other buffers known tothose of skill in the art. In one embodiment, the buffer is citricacid/phosphate buffer, acetate buffer, citrate buffer or phosphatebuffer. In another embodiment, the buffer is a citrate buffer (citricacid/sodium citrate). The buffer concentration has been found to affectthe stability of the composition. Buffer concentrations for use includefrom about 0 or 0.01 mM to about 150 mM, or 0 or 0.01 mM to 150 mM. Inanother embodiment, the buffer concentration is about 1 mM to about 20mM, or 1 mM to 20 mM. In one embodiment, the buffer concentration isabout 5 mM, or 5 mM. In other embodiments, the buffer concentration isabout 1 mM to about 50 mM, or 1 mM to 50 mM. In one embodiment, thebuffer concentration is about 20 mM, or 20 mM. The kinetic-pH profile offormoterol is dependent on buffer concentration. At low andapproximately neutral conditions, increasing the buffer concentrationfrom 5 mM to 20 mM increased the rate constant of decompositionsignificantly. However, no noticeable differences in rate constant wereobserved in the pH region of about 4.5 to about 5.5, with increasingbuffer concentration from 5 mM to 20 mM. The particular buffer andbuffer concentration of a given composition for long term storageprovided herein may be determined empirically using standard stabilityassays well known to those of skill in the art (see, e.g., theExamples).

The ionic strength of the compositions provided herein also has beenfound to affect the stability of the composition. Ionic strengths of thecompositions provided herein are from about 0 to about 0.4, or 0 to 0.4.In another embodiment, the ionic strength of the compositions providedis about 0.05 to about 0.16, or 0.05 to 0.16. Compositions having alower ionic strength exhibit improved stability over formulations havinghigher ionic strength. The rate constant of decomposition wasessentially the same at ionic strength 0.05 to 0.1, but increased tosome extent at ionic strength of 0.2. The particular ionic strength of agiven composition for long term storage provided herein may bedetermined empirically using standard stability assays well known tothose of skill in the art (see, e.g., the Examples).

In embodiments where the pharmacologically suitable fluid is a salinesolution, tonicity adjusting agents may be added to provide the desiredionic strength. Tonicity adjusting agents for use herein include thosewhich display no or only negligible pharmacological activity afteradministration. Both inorganic and organic tonicity adjusting agents maybe used in the compositions provided herein. Tonicity adjusting agentsinclude, but are not limited to, ammonium carbonate, ammonium chloride,ammonium lactate, ammonium nitrate, ammonium phosphate, ammoniumsulfate, ascorbic acid, bismuth sodium tartrate, boric acid, calciumchloride, calcium disodium edetate, calcium gluconate, calcium lactate,citric acid, dextrose, diethanolamine, dimethylsulfoxide, edetatedisodium, edetate trisodium monohydrate, fluorescein sodium, fructose,galactose, glycerin, lactic acid, lactose, magnesium chloride, magnesiumsulfate, mannitol, polyethylene glycol, potassium acetate, potassiumchlorate, potassium chloride, potassium iodide, potassium nitrate,potassium phosphate, potassium sulfate, propylene glycol, silvernitrate, sodium acetate, sodium bicarbonate, sodium biphosphate, sodiumbisulfite, sodium borate, sodium bromide, sodium cacodylate, sodiumcarbonate, sodium chloride, sodium citrate, sodium iodide, sodiumlactate, sodium metabisulfite, sodium nitrate, sodium nitrite, sodiumphosphate, sodium propionate, sodium succinate, sodium sulfate, sodiumsulfite, sodium tartrate, sodium thiosulfate, sorbitol, sucrose,tartaric acid, triethanolamine, urea, urethan, uridine and zinc sulfate.In certain embodiments, the tonicity adjusting agent is sodium chloride.In these embodiments, the pharmacologically suitable fluid is aqueoussaline.

The storage temperature of the compositions provided herein also hasbeen found to affect the stability of the composition. Compositionsstored at a lower temperature exhibit improved stability overformulations stored at higher temperatures. The effect of temperature onthe rate constant of decomposition at pH 5, a buffer concentration of 5mM, and an ionic strength of 0.05, was linear according to Arrheniuskinetics, i.e., when Ln k_(obs) was plotted against 1/T, where T is thetemperature in degree Kelvin.

The estimated shelf-life of formoterol in the compositions providedherein is significantly greater than that reported for known formoterolcompositions. The estimated shelf-life of formoterol in the compositionsprovided herein is about 6.2 years, at 5° C. and about 7.5 months, or at25° C. The estimated formoterol concentrations in the compositionsprovided herein as a function of storage time at 5° C. and usage time at25° C. was determined. It is estimated that greater than 90% of theinitial formoterol present in the composition remains after 3 months ofusage time at 25° C. and 3 years of storage time at 5° C. as well asafter 0.5 months of usage time at 25° C. and 1 year of storage time at5° C.

In one embodiment, the compositions provided herein are preparedcontaining formoterol fumarate at a nominal concentration of 0.1 mg/mLat the indicated pH and citric acid/phosphate buffer concentrations. Thesolutions were stored at 60° C. In these compositions, formoterol isrelatively more stable at a pH from about 4 to about 5, and is also morestable at lower buffer concentration.

The compositions provided herein also may include excipients andadditives. The particular excipient or additive for use in thecompositions for long term storage provided herein may be determinedempirically using methods well known to those of skill in the art (see,e.g., the Examples). Excipients and additives are any pharmacologicallysuitable and therapeutically useful substance which is not an activesubstance. Excipients and additives generally have no pharmacologicalactivity, or at least no undesirable pharmacological activity. Theexcipients and additives include, but are not limited to, surfactants,stabilizers, complexing agents, antioxidants, or preservatives whichprolong the duration of use of the finished pharmaceutical formulation,flavorings, vitamins, or other additives known in the art. Complexingagents include, but are not limited to, ethylenediaminetetraacetic acid(EDTA) or a salt thereof, such as the disodium salt, citric acid,nitrilotriacetic acid and the salts thereof. In one embodiment, thecomplexing agent is EDTA. Preservatives include, but are not limited to,those that protect the solution from contamination with pathogenicparticles, including benzalkonium chloride or benzoic acid, or benzoatessuch as sodium benzoate. Antioxidants include, but are not limited to,vitamins, provitaimins, ascorbic acid, vitamin E or salts or estersthereof.

The compositions provided herein also may include a cosolvent, whichincreases the solubility of additives or the active ingredient(s). Theparticular cosolvent for use in the compositions for long term storageprovided herein may be determined empirically using methods well knownto those of skill in the art. Cosolvents for use herein include, but arenot limited to, hydroxylated solvents or other polar solvents, such asalcohols such as isopropyl alcohol, glycols such as propylene glycol,polyethylene glycol, polypropylene glycol, glycol ether, glycerol, andpolyoxyethylene alcohols.

C. Preparation of Compounds for Use in the Compositions

The preparation of the compounds used in the compositions providedherein is described below. Any such compound or similar compound may besynthesized according to a method discussed in general below or by onlyminor modification of the methods by selecting appropriate startingmaterials.

Formoterol may be prepared according to the method disclosed in U.S.Pat. No. 3,994,974. Briefly, 4-benzyloxy-3-nitro-a-bromoacetophenone isreacted with N-benzyl-N-(1-methyl-2-p-methoxyphenylethyl)amine to formthe α-aminoacetophenone. This compound was subjected to the followingseries of reactions: (i) reduction of the ketone with sodiumborohydride; (ii) reduction of the nitro group with aqueous hydrochloricacid and iron powder; (iii) amine formylation with acetic anhydride andformic acid; and (iv) catalytic reduction over 10% palladium on carbonto afford formoterol free base. Crystallization of the 12 fumarate saltfrom ethanol provides (formoterol)-½ fumarate.

The individual enantiomers of formoterol,2-hydroxy-5-((1S)-1-hydroxy-2-(((1S)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilideand2-hydroxy-5-((1R)-1-hydroxy-2-(((1R)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)formanilide,may be prepared by the method disclosed in U.S. Pat. No. 6,040,344.Briefly, reaction of optically pure 4-benzyloxy-3-formamidostyrene oxidewith an optically pure4-methoxy-a-methyl-N-(phenylmethyl)benzene-ethanamine, followed bydebenzylation, affords the desired enantiomer of formoterol.Debenzylation may be accomplished by reduction with hydrogen gas in thepresence of a noble metal catalyst, such as palladium on carbon.

The required optically pure 4-benzyloxy-3-formamidostyrene oxide may beprepared from 4-benzyloxy-3-nitro-a-bromoacetophenone by (i) reductionwith vorane in the presence of an optically pure aminoindanol, (ii)hydrogenation over platinum oxide catalyst, (iii) formylation withformic acid and acetic anhydride, and (iv) epoxide formation in thepresence of potassium carbonate.

The required optically pure4-methoxy-a-methyl-N-(phenylmethyl-1)benzeneethanamine may be preparedfrom 4-methoxyphenylacetone by (i) reductive amination with benzylaminein the presence of hydrogen and a platinum catalyst, and (ii)crystallization of the desired optically pure amine from the resultingracemic mixture as its mandelic acid salt.

D. Formulation of Pharmaceutical Compositions

The compositions provided herein are prepared by procedures well knownto those of skill in the art. For example, a formoterol fumaratesolution may be prepared by the procedure of EXAMPLE 1. Briefly, abuffer solution having a pH and ionic strength of interest herein isprepared. In one embodiment, the buffer is a mixture of citric acid andsodium citrate, with sodium chloride added to achieve the desired ionicstrength. Formoterol fumarate dihydrate is added to the buffer solutionwith agitation to produce a solution of the desired formoterolconcentration. Exemplary formoterol concentrations is 0.0021 kgformoterol fumarate dihydrate/100 kg water.

E. Evaluation of the Activity of the Compositions

Standard physiological, pharmacological and biochemical procedures areavailable for testing the compositions provided herein to identify thosethat possess bronchodilatory activity.

In vitro and in vivo assays that may be used to evaluate bronchodilatoryactivity are well known to those of skill in the art. See also, e.g.,U.S. Pat. Nos. 3,994,974, and 6,068,833; German Patent No. 2,305,092;Kaumann et al. (1985) Naunyn-Schmied Arch. Pharmacol. 331:27-39; Lemoineet al. (1985) Naunyn-Schmied Arch. Pharmacol. 331:40-51; Tomioka et al.(1981) Arch. Int. Pharmacodyn. 250:279-292; Dellamary et al. (2000)Pharm. Res. 17(2):168-174; Rico-Mendez et al. (1999) Rev. Alerg. Mex.46(5):130-135; Seberova et al. (20.00) Respir. Med. 94(6):607-611;Lotvall et ald (1999) Can. Respir. J. 6(5):412-416; Campbell et al.(1999) Respir. Med. 93(4):236-244; Nightingale et al. (1999) Am. J.Respir. Crit. Care Med. 159(6):1786-1790; Lecaillon et al. (1999) Eur.J. Clin. Pharmacol. 55(2):131-138; Bartow et al. (1998) Drugs55(2):303-322; Ekstrom et al. (1998) Respir. Med. 92(8):1040-1045;Ringdal et al. (1998) Respir. Med. 92(8):1017-1021; Totterman et al.(1998) Eur. Respir. J. 12(3):573-579; Palmqvist et al. (1997) Eur.Respir. J. 10 (11):2484-2489; Nielsen et al. (1997) Eur. Respir. J.10(9):2105-2109; Ullman et al. (1996) Allergy 51(10):745-748; Selroos etal. (1996) Clin. Immunother. 6:273-299; and Schreurs et al. (1996) Eur.Respir. J. 9(8):1678-1683.

F. Methods of Treatment of Bronchoconstrictive Disorders

The compositions provided herein are used for treating, preventing, orameliorating one or more symptoms of a bronchoconstrictive disorders ina subject. En one embodiment, the method includes administering to asubject an effective amount of a composition containing abronchodilating agent, including, but not limited to, formoterol,whereby the disease or disorder is treated or prevented. The subjecttreated is, in certain embodiments, a mammal. The mammal treated is, incertain embodiments, a human.

In another embodiment, the method provided herein includes oraladministration of a composition provided herein. In certain embodimentsherein, the composition is directly administered to a subject in need ofsuch treatment via nebulization without dilution or other modificationof the composition prior to administration.

The methods for treatment, prevention, or amelioration of one or moresymptoms of bronchoconstrictive disorders, in another embodiment,further include administering one or more of (a), (b), (c) or (d) asfollows: (a) a β₂-adrenoreceptor agonist; (b) a dopamine (D₂) receptoragonist; (c) a prophylactic therapeutic, such as a steroid; or (d) ananticholinergic agent; simultaneously with, prior to or subsequent tothe composition provided herein.

β₂-Adrenoreceptor agonists for use in combination with the compositionsprovided herein include, but are not limited to, Albuterol(a¹-(((1,1-dimethylethyl)amino)methyl)-4-hydroxy-1,3-benzenedimethanol);Bambuterol (dimethylcarbamic acid5-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,3-phenylene ester);Bitolterol (4-methylbenzoic acid4-(2-(1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,2-phenylene ester);Broxaterol(3-bromo-a-(((1,1-dimethylethyl)amino)methyl)-5-isoxazolemethanol);Isoproterenol(4-(1-hydroxy-2-((1-methylethyl)amino)ethyl)-1,2-benzenediol);Trimetoquinol(1,2,3,4-tetrahydro-1-((3,4,5-trimethoxyphenyl)methyl)-6,7-isoquinolinediol),Clenbuterol(4-amino-3,5-dichloro-a-(((1,1-dimethylethyl)amino)methyl)benzenemethanol);Fenoterol(5-(1-hydroxy-2-((2-(4-hydroxyphenyl)-1-methylethyl)amino)ethyl)-1,3-benzenediol);Formoterol(2-hydroxy-5-((1RS)-1-hydroxy-2-(((1RS)-2-(p-methoxyphenyl)-1-methylethyl)amino)ethyl)-formanilide);(R,R)-Formoterol; Desformoterol ((R,R) or(S,S)-3-amino-4-hydroxy-a-(((2-(4-methoxyphenyl)-1-methylethyl)amino)methyl)benzenemethanol);Hexoprenaline(4,4′-(1,6-hexanediyl)-bis(imino(1-hydroxy-2,1-ethanediyl)))bis-1,2-benzenediol);Isoetharine(4-(1-hydroxy-2-((1-methylethyl)amino)butyl)-1,2-benzenediol);Isoprenaline(4-(1-hydroxy-2-((1-methylethyl)-amino)ethyl)-1,2-benzenediol);Metaproterenol(5-(1-hydroxy-2-((1-methylethyl)amino)ethyl)-1,3-benzene-diol);Picumeterol(4-amino-3,5-dichloro-a-(((6-(2-(2-pyridiyl)ethoxy)hexyl)-amino)methyl)benzenemethanol);Pirbuterol(a⁶-(((1,1-dimethylethyl)amino)methyl)-3-hydroxy-2,6-pyridinemethanol);Procaterol((R*,S*)-(±)-8-hydroxy-5-(1-hydroxy-2-((1-methylethyl)amino)butyl)-2(-1H)-quinolinone);Reproterol((7-(3-((2-(3,5-dihydroxyphenyl)-2-hydroxyethyl)amino)-propyl)-3,7-dihydro-1,3-dimethyl-1H-purine-2,6-dione);Rimiterol (4-(hydroxy-2-piperidinylmethyl)-1,2-benzenediol); Salbutamol((±)-a¹-(((1,1-dimethylethyl)-amino)-methyl)-4-hydroxy-1,3-benzenedinethanol);(R)-Salbutamol; Salmeterol((±)-4-hydroxy-a¹-(((6-(4-phenylbutoxy)hexyl)amino)methyl)-1,3-benzenedimethanol);(R)-Salmeterol; Terbutaline(5-(2-((1,1-dimethylethyl)amino)-1-hydroxyethyl)-1,3-benzenediol);Tulobuterol(2-chloro-a-(((1,1-dimethylethyl)amino)-methyl)benzenemethanol); andTA-2005(8-hydroxy-5-((1R)-1-hydroxy-2-(N-((1R)-2-(4-methoxyphenyl)-1-methylethyl)amino)ethyl)-carbostyrilhydrochloride).

Dopamine (D₂) receptor agonists include, but are not limited to,Apomorphine((r)-5,6,6a,7-tetrahydro-6-methyl-4H-dibenzo[de,g]quinoline-10,11-diol);Bromocriptine ((5′a)-2-bromo-12′-hydroxy-2′-(1-methylethyl)-5′-(2-methylpropyl)ergotaman-3′,6′,18-trione);Cabergoline((8β)-N-(3-(dimethylamino)propyl)-N-((ethylamino)carbonyl)-6-(2-prop-enyl)ergoline-8-carboxamide);Lisuride (N′-((8a)-9,10-didehydro-6-methylergolin-8-yl)N,N-diethylurea);Pergolide ((8a)-8-((methylthio)methyl)-6-propylergoline); Levodopa(3-hydroxy-L-tryrosine); Pramipexole((s)-4,5,6,7-tetrahydro-N.sup.6-propyl-2,6-benzothiazolediamine);Quinpirole hydrochlrodie(trans-(−)-4aR-4,4-a,5,6,7,8,8a,9-octahydro-5-propyl-1H-pyrazolo[3,4-g]quinolinehydrochloride); Ropinirole(4-(2-(dipropylamino)ethyl)-1,3-dihydro-2H-indol-2-one); and Talipexole(5,6,7,8-tetrahydro-6-(2-propenyl)-4H-thiazolo[4,5-d]azepin-2-amine).Other dopamine D₂ receptor agonists for use herein are disclosed inInternational Patent Application Publication No. WO 99/36095.

Prophylactic therapeutics for use in combination therapy herein includesteroidal anti-inflammatory agents, including, but not limited to,beclomethasone dipropionate (BDP), beclomethasone monopropionate (BMP),flunisolide, triamcinolone acetonide, dexamethasone, tipredane,ciclesonid, rofleponide, mometasone, mometasone fliroate (ASMANEX®TWISTHALER™, Schering-Plough Corporation, Kenilworth, N.J.), RPR 106541,having the formula

fluticasone or fluticasone propionate and budesonide or by way of sodiumcromoglycate or nedocromil sodium.

Anticholinergic agents for use herein include, but are not limited to,ipratropium bromide, oxitropium bromide, atropine methyl nitrate,atropine sulfate, ipratropium, belladonna extract, scopolamine,scopolamine methobromide, homatropine methobromide, hyoscyamine,isopriopramide, orphenadrine, benzalkonium chloride, tiotropium bromideand glycopyrronium bromide. In certain embodiments, the compositionscontain an anticholinergic agent, such as ipratropium bromide, at aconcentration of about 100 μg/mL to about 500 μg/mL, or 100 μg/mL to 500μg/mL. In other embodiments, ipratropium bromide concentration is about150 μg/mL to about 350 μg/mL, or 150 μg/mL to 350 μg/mL. In otherembodiments, the compositions for use in the methods herein containipratropium bromide at a concentration of about 200 μg/mL to about 300μg/ml, or 200 μg/ml to 300 μg/mL. In other embodiments, the compositionsfor use in the methods herein contain ipratropium bromide at aconcentration of about 250 μg/mL, or 250 μg/mL.

Other active ingredients for use herein in combination therapy, include,but are not limited to, IL-5 inhibitors such as those disclosed in U.S.Pat. Nos. 5,668,110, 5,683,983, 5,677,280 and 5,654,276; antisensemodulators of IL-5 such as those disclosed in U.S. Pat. No. 6,136,603;milrinone (1,6-dihydro-2-methyl-6-oxo-[3,4′-bipyridine]-5-carbonitrilemilrinone lactate; tryptase inhibitors such as those disclosed in U.S.Pat. No. 5,525,623; tachykinin receptor antagonists such as thosedisclosed in U.S. Pat. Nos. 5,691,336, 5,877,191, 5,929,094, 5,750,549and 5,780,467; leukotriene receptor antagonists such as montelukastsodium (SINGULAR®,R-(E)]-1-[[[1-[3-[2-(7-chloro-2-quinolinyl)ethenyl-]phenyl]-3-[2-(1-hydroxy-1-methylethyl)-phenyl]-propyl]thio]methyl]cyclopto-paneaceticacid, monosodium salt), 5-lypoxygenase inhibitors such as zileuton(ZYFLO®, Abbott Laboratories, Abbott Park, Ill.), and anti-IgEantibodies such as XOLAIR® (recombinant humanized anti-IgE monoclonalantibody (CGP 51901; IGE 025A; rhuMAb-E25), Genentech, Inc., South SanFrancisco, Calif.).

The bronchoconstrictive disorder to be treated, prevented, or whose oneor more symptoms are to be ameliorated is associated with asthma,including, but not limited to, bronchial asthma, allergic asthma andintrinsic asthma, e.g., late asthma and airway hyper-responsiveness;and, particularly in embodiments where an, anticholinergic agent isused, other chronic obstructive pulmonary diseases (COPDs), including,but not limited to, chronic bronchitis, emphysema, and associated corpulmonale (heart disease secondary to disease of the lungs andrespiratory system) with pulmonary hypertension, right ventricularhypertrophy and right heart failure. COPD is frequently associated withcigarette smoking, infections, environmental pollution and occupationaldust exposure.

G. Nebulizers

The compositions provided herein are intended for administration to asubject in need of such treatment via nebulization. Nebulizers thatnebulize liquid formulations containing no propellant are suitable foruse with the compositions provided herein. The nebilizer and can be unitdose or multidose. Nebulizers are available from, e.g., Pari GmbH(Stamberg, Germany), DeVilbiss Healthcare (Heston, Middlesex, UK),Healthdyne, Vital Signs, Baxter, Allied Health Care, Invacare, Hudson,Omron, Bremed, AirSep, Luminscope, Medisana, Siemens, Aerogen, MountainMedical, Aerosol Medical Ltd. (Colchester, Essex, UK), AFP Medical(Rugby, Warwickshire, UK), Bard Ltd. (Sunderland, UK), Carri-Med Ltd(Dorking, UK), Plaem Nuiva (Brescia, Italy), Henleys Medical Supplies(London, UK), Intersurgical (Berkshire, UK), Lifecare Hospital Supplies(Leies, UK), Medic-Aid Ltd. (West Sussex, UK), Medix Ltd. (Essex, UK),Sinclair Medical Ltd. (Surrey, UK), and many others.

Nebulizers for use herein include, but are not limited to, jetnebulizers (optionally sold with compressors), ultrasonic nebulizers,and others. Exemplary jet nebulizers for use herein include Pari LCplus/ProNeb, Pari LC plus/ProNeb Turbo, Pari LC plus/Dura Neb 1000 &2000, Pari LC plus/Walkhaler, Pari LC plus/Pari Master, Pari LC star,Omron CompAir XL Portable Nebulizer System (NE-C18 and JetAir Disposablenebulizer), Omron CompAir Elite Compressor Nebulizer System (NE-C21 andElite Air Reusable Nebilizer), Pari LC Plus or Pari LC Star nebulizerwith Proneb Ultra compressor, Pulmo-aide, Pulmo-aide LT, Pulmo-aidetraveler, Invacare Passport, Inspiration Healthdyne 626, Pulmo-NebTraveler, DeVilbiss 646, Whisper Jet, Acorn II, Misty-Neb, Alliedaerosol, Schuco Home Care, Lexan Plasic Pocet Neb, SideStream Hand HeldNeb, Mobil Mist, Up-Draft, Up-Draft II, T Up-Draft, ISO-NEB, AVA-NEB,Micro Mist, and PulmoMate. Exemplary ultrasonic nebulizers for useherein include MicroAir, Ultai, Siemens Ultra Nebulizer 145, CompAir,Puinosonic, Scout, 5003 Ultrasonic Neb, 5110 Ultrasonic Neb, 5004 DeskUltrasonic Nebulizer, Mystique Ultrasonic, Luminscope's UltrasonicNebulizer, Medisana Ultrasonic Nebulizer, Microstat UltrasonicNebulizer, and MABISMist Hand Held Ultrasonic Nebulizer. Othernebulizers for use herein include 5000 Electromagnetic Neb, 5001Electromagnetic Neb 5002 Rotary Piston Neb, Lumineb I Piston Nebulizer5500, AERONEB™ Portable Nebuizer System, AERODOSE™ Inhaler, AeroEclipseBreath Actuated Nebulizer, HALOLITE™ system (Profile Therapeutics),AKITA® systems (InaMed, Germany), Mystic system (BattellePharma),RESPIMAT® (Boehringer Ingelheim), AERX® (Aradigm), and E-FLOW™ (Pari).

Depending on the nebulizer used, the volume of the formoterol inhalationsolution nebulized in one embodiment is about 0.1 mL to 3 mL, or 0.1 mLto 3 mL. In another embodiment, the volume is about 2 mL, or 2 mL. Inanother embodiment, the volume is about 1 mL, or 1 mL. In anotherembodiment, the volume is about 0.5 mL, or 0.5 mL.

H. Articles of Manufacture

The compositions provided herein may be packaged as articles ofmanufacture containing packaging material, a composition providedherein, which is useful for treatment, prevention or amelioration of oneor more symptoms of diseases or disorders associated with undesiredand/or uncontrolled bronchoconstriction, and a label that indicates thatthe composition is used for treatment, prevention or amelioration of oneor more symptoms of diseases or disorders associated with undesiredand/or uncontrolled bronchoconstriction.

The articles of manufacture provided herein contain packaging materials.Packaging materials for use in packaging pharmaceutical products arewell known to those of skill in the art. See, e.g., U.S. Pat. Nos.5,323,907, 5,052,558 and 5,033,252. Examples of pharmaceutical packagingmaterials include, but are not limited to, blister packs, bottles,tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, andany packaging material suitable for a selected formulation and intendedmode of administration and treatment.

In one embodiment herein, the compositions are packaged with a nebulizerfor direct administration of the composition to a subject in needthereof.

The following examples are included for illustrative purposes only andare not intended to limit the scope of the invention.

EXAMPLE 1 Preparation of Formoterol Inhalation Solution Formulation

Appropriate quantities of the raw materials are weighed for the 100 Kgbatch as shown below.

20 μg/mL* 10 μg/mL* Formoterol fumarate dihydrate 0.0021 kg 0.00105 kgCitric acid monohydrate USP 0.135 kg 0.135 kg Sodium Citrate dihydrateUSP 0.400 kg 0.400 kg Sodium chloride USP 0.785 kg 0.785 kg Purifiedwater USP q.s. to 100 kg q.s. to 100 kg *Concentration of formoterolfumarate (anhydrous)

In a clean stainless steel (SS) tank fitted with bottom drain, 75% ofthe required amount of purified water is added. Samples are taken forpH, conductivity, and microbiological testing. Citric acid monohydrate,sodium citrate dihydrate and sodium chloride are added to the tank andmixed for 15 minutes to dissolve. A sample is taken at this point tocheck pH. Formoterol fumarate dihydrate is added at this point and mixedfor about 75 minutes to dissolve all active raw material. Purified wateris used to adjust to final volume. The formulation is mixed for anadditional 30 minutes and samples for pH and assay are taken based onwhich the formulation is released for filling. The bulk solution isfilled into low density polyethylene (LDPE) vials (2 mL fill) in aform-fill-seal (FFS) machine. The released drug product solution istransferred from the formulation tank through sanitary delivery linesinto the FFS machine. The individual vials are overwrapped with asuitable foil laminate.

EXAMPLE 2 Procedure for Stability Testing of Formoterol Solutions

Stability samples of the solution prepared in EXAMPLE 1 and solution offormoterol fumarate (20 μg/mL) and ipratropium bromide (250 μg/mL) wereplaced in LDPE vials and stored in stability ovens at acceleratedtemperatures. At selected time points, aliquots of the samples wereremoved from the vials. The formoterol concentrations of the sampleswere analyzed by high performance liquid chromatography.

Provided herein is the stability data for exemplary formulationscontaining formoterol and formoterol in combination with ipratropiumbromide.

Stability data on formoterol (20 μg/mL) and formoterolfumarate/ipratropium bromide combination (20 μg/ml and 250 μg/mL):

Assay as percent of label claim Formoterol fumarate/ Formterolipratropium bromide Inhalation solution inhalation solution Storagecondition Formoterol Formoterol Ipratropium Initial 100 100.5 101.2  5°C./3 months 96.7 100 101.6 25° C./3 months 94.5 100 101.2

Since modifications will be apparent to those of skill in this art, itis intended that this invention be limited only by the scope of theappended claims.

1. A method of treating undesired and/or uncontrolledbronchoconstriction comprising the steps of: (a) adding to a nebulizer,from a propellant-free, sterile unit dose package about 0.1 to about 3.0mL of a pharmaceutical composition comprising (R) formoterol or a saltthereof at a concentration of from about 0.08 μg/mL to about 43 μg/mLbased on formoterol free base, in a pharmacologically suitable solution,wherein the composition further comprises water and a buffer having aconcentration of from about 1 mM to about 50 mM, said composition havinga pH of about 4.0 to about 6.0, and having an estimated shelf life ofgreater than 90% after 3 months storage at 25° C. and after 3 yearsstorage at 5° C. (b) directly administering said composition to asubject in need thereof, without dilution or other modification of saidprior to administration.
 2. A method of treating undesired and/oruncontrolled bronchoconstriction comprising the steps of: (a) adding toa nebulizer from a propellant-free, sterile unit dose package about 0.1to about 3.0 mL of a pharmaceutical composition comprising (R)formoterol or a salt thereof at a concentration of from about 0.08 μg/mLto about 43 μg/mL based on formoterol free base, in a pharmacologicallysuitable solution, wherein the composition further comprises water and abuffer having a concentration of from about 1 mM to about 50 mM, saidcomposition having a pH of about 4.0 to about 6.0, and having anestimated shelf life of greater than about 94% after 3 months storage at25° C. and greater than about 96% after 3 months storage at 5° C.; and(b) directly administering said composition to a subject in needthereof, without dilution or other modification of said prior toadministration.
 3. A method of treating undesired and/or uncontrolledbronchoconstriction comprising the steps of: (a) adding from apropellant-free, sterile unit dose package to a nebulizer about 0.1 toabout 3.0 mL of a pharmaceutical composition comprising (E) formoterolor a salt thereof at a concentration of from about 0.08 μg/mL to about43 μg/mL based on formoterol free base, in a pharmacologically suitablesolution, wherein the composition further comprises water and a citratebuffer at a concentration of from about 1 mM to about 20 mM, saidcomposition having a pH of about 4.5 to about 5.5; and (b) directlyadministering said composition to a subject in need thereof, withoutdilution or other modification of said composition prior toadministration.
 4. The method of treating undesired and/or uncontrolledbronchoconstriction as in any one of claims 1, 2 and 3 wherein saidundesired and/or uncontrolled bronchoconstriction is a symptom orcondition of asthma.
 5. The method of treating undesired and/oruncontrolled bronchoconstriction as in any one of claims 1, 2 and 3wherein said undesired and/or uncontrolled bronchoconstriction is asymptom or condition of chronic obstructive pulmonary disease.
 6. Themethod of treating undesired and/or uncontrolled bronchoconstriction asin any one of claims 1, 2 and 3 wherein said buffer is present at aconcentration of between about 1 mM and about 50 mM.
 7. The method oftreating undesired and/or uncontrolled bronchoconstriction as in any oneof claims 1 and 2 wherein said buffer is present at a concentration ofbetween about 1 mM and about 20 mM.
 8. The method of treating undesiredand/or uncontrolled bronchoconstriction as in any one of claims 1, 2 and3 wherein said composition has a pH of about
 5. 9. The method oftreating undesired and/or uncontrolled bronchoconstriction of claim 8wherein said buffer has a pH of about
 5. 10. The method of treatingundesired and/or uncontrolled bronchoconstriction as in any one ofclaims 1 and 2, wherein about 2.0 mL of the composition is nebulized.11. The method of treating undesired and/or uncontrolledbronchoconstriction as in any of claims 1, 2 or 3, wherein said (R)formoterol or a salt thereof is (R) formoterol tartrate.